| 大肠杆菌莽草酸途径限速酶多基因盒的构建及基因替换 |
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| 引用本文： | 李永辉,王世春,刘云,郭长江,蒋与刚,徐琪寿. 大肠杆菌莽草酸途径限速酶多基因盒的构建及基因替换[J]. 中国生物化学与分子生物学报, 2004, 20(4): 473-478 |
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| 作者姓名： | 李永辉 王世春 刘云 郭长江 蒋与刚 徐琪寿 |
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| 作者单位： | 1. 军事医学科学院放射医学研究所,北京,100850
2. 军事医学科学院卫生学与环境医学研究所,天津,300050 |
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| 基金项目： | 军事医学科学院创新项目，天津市科技攻关项目 |
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| 摘 要： | 优化大肠杆菌芳香族氨基酸生物合成代谢途径 ,构建莽草酸代谢途径限速酶的多基因盒PtacaroAaroCaroBkan .利用Red重组系统 ,在破坏整体调控基因csrA时 ,替换多基因盒 .Southern印迹证实 ,基因破坏和基因替换是成功的 .摇瓶发酵表明 ,构建的基因工程菌株比原始菌株基础产酸率提高了 4 5 3倍
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| 关 键 词： | 多基因盒子 Red重组系统 基因破坏和基因替换 |
| 收稿时间： | 2004-08-20 |
| 修稿时间： | 2003-08-15 |
Construction of Rate-Limiting Multigene-Cassette in Shikimate Pathway and Gene Replacement in E.coli |
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| LI Yong hui ),WANG Shi chun ),LIU Yun ),Guo Chang jiang ),Jiang Yu gang ),XU Qi shou ). Construction of Rate-Limiting Multigene-Cassette in Shikimate Pathway and Gene Replacement in E.coli[J]. Chinese Journal of Biochemistry and Molecular Biology, 2004, 20(4): 473-478 |
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| Authors: | LI Yong hui ) WANG Shi chun ) LIU Yun ) Guo Chang jiang ) Jiang Yu gang ) XU Qi shou ) |
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| Affiliation: | ( 1) Institute of Radiation Medicine, Academy of Military Medicine Sciences,Beijing 100850, China; 2) Institute of Hygiene and Environmental Medicine, Academy of Military Medicine Sciences, Tianjin 300050 China |
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| Abstract: | Multigene cassette carrying Ptac, aroA, aroB, aroC and kan was constructed to optimize aromatic amino acids biosynthesis pathway in E.coli. The global regulatory gene csrA was disrupted and replaced by multigene cassette using Red recombinant system. Southern blotting confirmed that the gene disruption and replacement were successful. The flasks fermentation verified that the rate of conversion from glucose to amino acid of the modified strain increased 4.53 folds than that of the control. |
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| Keywords: | multigene cassette Red recombinant system gene disruption and replacement |
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